Background and Objective; Ang-1 has implication on angiogenesis with reduced vascular leakage compared to VEGF and enhancing endothelial cell survival. COMP-Ang1 is a potent variant of Ang-1 to overcome insolubility, biological instability of Ang-1. There are very scarce data of COMP-Ang1 effect on jeopardized cardiomyocyte (CMC) with oxidative stress, especially in the aspect of cell survival signaling. Therefore, we examined the role of COMP-Ang1 against oxidative stress induced by H²O² in rat CMCs (rCMCs) via cell survival signal.
Methods; Primary cultures of neonatal rat CMCs were prepared from the ventricles of 1-day-old Sprague-Dawley rats. 5mM of COMP-Ang1 or media as control was pretreated to CMC (3X10⁵) before the oxidative stress. Oxidative stress was induced by 0.1mM of H²O² for 2 hours in 2 groups. Apoptosis severity was measured by FACS analysis with Annexin-���-FITC staining. And also, protein related with cell survival pathway and apoptosis (p-Akt/Akt, p-p38/p38, Bcl-2/Bax, NF-kB/IkB, Mn-SOD and caspase-3) were measured by Western blot.
Results;COMP-Ang1 effectively reduced H²O² related CMC apoptosis compared to control (14.6% vs 27.76%, difference 90.1%). Also, the expressions of p-Akt /Akt, Bcl-2/Bax, Mn-SOD were significantly increased in COMP-Ang1 group (OD result in Table2). However Caspase-3, p38/p-p38 and NF-kB/IkB, as the executors of apoptosis signal, was decreased in COMP-Ang1 group.
Conclusion;COMP-Ang1 protects CMC from apoptosis in oxidative stress generated with H2O2 via cell survival molecule overexpression.